摘要
目的从法尼醇受体(farnesoid X receptor,FXR)-硫酸基转移酶1E1(sulfotransferase 1E1,SULT1E1)通路探讨盐补骨脂致小鼠胆汁淤积性肝损伤性别差异的潜在机制。方法ICR雌性和雄性小鼠连续ig不同剂量(5、10、20 g/kg)的盐补骨脂水煎液30 d,采用HPLC仪检测补骨脂炮制后化学成分含量;采用全自动生化仪检测肝损伤相关指标;取肝脏、脾脏、肾脏、肺脏以及卵巢、睾丸,称定质量并计算脏器指数;采用苏木素-伊红(HE)染色法进行肝脏病理组织学检查;采用透射电镜观察盐补骨脂致胆汁淤积性肝损伤的显微结构变化;采用ELISA法检测血清中雌二醇(estradiol,E_(2))、促卵泡激素(follicle-stimulating hormone,FSH)、促黄体生成素(luteinizing hormone,LH)、白细胞介素-6(interleukin-6,IL-6)、对氧磷酶1(paraoxonase 1,PON1)及FXR含量;采用免疫荧光染色法检测肝脏胆盐输出泵(bile salt export pump,Bsep)、多药耐药相关蛋白2(multidrug resistance protein 2,Mrp2)和SULT1E1的表达;采用Western blotting检测肝脏FXR、Mrp2、胆固醇7α-羟化酶(cholesterol 7α-hydroxylase,CYP7a1)、Bsep、SULT1E1、肝细胞核因子4α(hepatocyte nuclear factor 4α,HNF4α)和核因子E2相关因子2(nuclear factor erythroid-2-related factor 2,Nrf2)蛋白表达。结果盐补骨脂给药后小鼠肝脏出现明显的肝细胞肿胀、胆管扩张、血清生化指标升高等病理变化,其中雌鼠更为严重(P<0.05、0.01、0.001)。免疫荧光实验显示给药后肝脏Bsep和Mrp2荧光强度均显著降低(P<0.05、0.001),但雌鼠更为明显;雌鼠肝脏中SULT1E1表达水平显著降低(P<0.001),而在雄鼠的肝脏中没有差异。ELISA结果显示雌鼠和雄鼠血清中E2、FSH和LH水平均显著升高(P<0.05、0.01、0.001),雌鼠更为明显。Western blotting结果显示,雌鼠给药后肝脏FXR、Mrp2、Bsep、SULT1E1、HNF4α和Nrf2表达明显降低(P<0.05、0.01、0.001),CYP7a1表达显著升高(P<0.01);雄鼠给药后FXR和Bsep表达明显降低(P<0.
Objective To explore the potential mechanism of salt-processed Buguzhi(Psoraleae Fructus)induced gender difference of cholestatic liver injury in mice through farnesoid X receptor(FXR)-sulfotransferase 1E1(SULT1E1)pathway.Methods ICR female and male mice were subjected to continuous ig of different dosages(5,10,20 g/kg)of salt-processed Psoraleae Fructus water decoction for 30 d,and chemical composition content of processed Psoraleae Fructus was detected by HPLC;Fully automated biochemical analyzer was used to detect liver injury related indicators;Liver,spleen,kidney,lung,ovary and testis were taken and weighed,organ index was calculated.Hematoxylin-eosin(HE)staining method was used for histopathological examination of liver.The microscopic structure of cholestatic liver injury induced by salt-processed Psoraleae Fructus was observed by transmission electron microscopy.ELISA was used to detect the levels of estradiol(E_(2)),follicle stimulating hormone(FSH),luteinizing hormone(LH),interleukin-6(IL-6),paraoxonase 1(PON1)and FXR in serum;The expressions of bile salt export pump(Bsep),multiple drug resistance protein 2(Mrp2)and SULT1E1 in liver were detected by immunofluorescence staining;Western blotting was used to detect FXR,Mrp2,cholesterol 7α-hydroxylase(CYP7a1),Bsep,SULT1E1,hepatocyte nuclear factor 4α(HNF4α)and nuclear factor E2 related factor 2(Nrf2)protein expressions in liver.Results After the administration of salt-processed Psoraleae Fructus,liver of mice showed significant liver cell swelling,bile duct dilation,and elevated serum biochemical indicators,with female mice showing more severe pathological changes(P<0.05,0.01,0.001).The immunofluorescence experiment showed that fluorescence intensity of Bsep and Mrp2 in liver were significantly decreased after administration(P<0.05,0.001),but it was more pronounced in female mice;The expression level of SULT1E1 was significantly reduced in liver of female mice(P<0.001),while there was no difference in liver of male mice.ELISA results showed a significant
作者
吴育
周浓
梅春梅
石芸
华政颖
李伟东
WU Yu;ZHOU Nong;MEI Chun-mei;SHI Yun;HUA Zheng-ying;LI Wei-dong(Nantong Hospital of Traditional Chinese Medicine,Nanjing University of Chinese Medicine,Nantong 226000,China;Jiangsu Key Laboratory of Chinese Medicine Processing,Engineering Center of Normalization and Standardization of Chinese Medicine Processing of Ministry of Education,Nanjing University of Chinese Medicine,Nanjing 210023,China;Chongqing Three Gorges University,Chongqing 404120,China)
出处
《中草药》
CAS
CSCD
北大核心
2023年第9期2802-2811,共10页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金面上项目(81773902)
国家自然科学基金面上项目(81973484)
2022年南京中医药大学自然科学基金资助项目(XZR2021085)。
